Mitigation of haemato-genotoxic and stress response effects in Cyprinus carpio via silymarin dietary supplementation following deltamethrin exposure.

The study examined the potential of Silymarin, a blend of bioactive flavonolignans extracted from the milk thistle Silybum marianum, to mitigate Deltamethrin-induced toxicity in the blood of Cyprinus carpio. Fish were exposed to Deltamethrin (0.66 µg/L), the plant extract, or a combination of both for a duration of thirty days. Various parameters, including serum biochemical markers, erythrocytic abnormalities, and genotoxicity endpoints, were assessed. Results indicated a significant (p < 0.05) increase in the levels of AST, ALT, ALP, blood urea nitrogen, creatinine, glucose, cholesterol, and TLC in the fish exposed to the pesticide. Conversely, total protein, TEC, and Hb showed a notable decrease. There was also a notable rise in micronuclei and erythrocytic abnormalities such as acanthocytes, microcytes, and notched cells. Under ultrastructural examination, phenotypic deformities like spherocytosis, discocytes, and clumped erythrocytes were observed. However, dietary supplementation of silymarin (1 g/kg) significantly restored the biochemical, genetic, and cellular parameters, resembling those of the control group. This suggests the potential of this plant extract in protecting the common carp, Cyprinus carpio, from Deltamethrin-induced damage by scavenging free radicals and reducing DNA oxidative stress.

Mitigating the nephrotoxic impact of hexavalent chromium in Ctenopharyngodon idellus (grass carp) with Boerhavia diffusa (punarnava) leaf extract.

In Ctenopharyngodon idellus, the ameliorative influence of rutin-containing leaf extract of Boerhavia diffusa was assessed against chronic exposure to hexavalent chromium. For this, alterations in chromium accumulation, oxidative stress, kidney function markers, histopathology (light and transmission electron microscopy), and transcriptional profiling (Nrf2 and MT2) were examined. RP-HPLC analysis confirmed the presence of rutin (90.45 ± 0.98 mg/g) in the ethanolic leaf extract of the plant. LD50 of the extract to the fish was beyond 5000 mg/kg b.w. The fish was subjected to a sublethal concentration of hexavalent chromium (5.30 mg/L) accompanied by a dose of 250 mg/kg b.w./day of extract in the diet for the experimental duration of 45 days. The extract alone did not generate any adverse consequences in the nephric tissue. Chronic exposure to hexavalent chromium damaged tissue irreparably, demonstrated by elevated levels of kidney function markers (blood urea nitrogen and creatinine) and altered histoarchitecture (DTC value of 78.02 ± 10.5). The metal exposure increased chromium accumulation and malondialdehyde (MDA) and decreased the reduced glutathione (GSH) levels, the activity of antioxidant enzymes (superoxide dismutase, catalase and glutathione-S-transferase) and gene expression in the tissue. The co-supplementation of leaf extract with metal exposure revealed a tissue architecture with normal to slight modifications, and the level of kidney markers, antioxidants, and genes expressed in a normalized range. Principal component analysis created two components with antioxidants (GSH, SOD, CAT, and GST) revealing a negative correlation with the second component comprising MDA, DTC, and chromium concentration. It can be concluded that B. diffusa leaves are safe additives in the fish diet and possess an ameliorative capacity for renal injury incurred by hexavalent chromium.

In vivo genotoxic effects of commercial grade cypermethrin on fish peripheral erythrocytes.

The study explicates the genotoxic effects of commercial grade cypermethrin on peripheral erythrocytes of Catla catla, chronically exposed to two environmentally relevant concentrations. The fish was treated with sub-lethal concentrations 0.12 µg/L and 0.41 µg/L (1/10th and /1/3rd of 96 h LC50 ) of cypermethrin for 45 days. DNA damage in the exposed fish was assessed using alkaline comet assay, presence of micronuclei (MN), erythrocyte nuclear and cytoplasmic abnormalities. Exposure to cypermethrin induced a dose-dependent increase in percent DNA damage, micronucleus frequency and erythrocyte abnormalities. Nuclear anomalies such as notched nuclei, lobed nuclei, bridged nuclei, and deformed nuclei; and cytoplasmic anomalies like anisochromasia, vacuolated cytoplasm, lobed cells, and echinocytes were observed. The findings revealed the genotoxic potential of commercial formulations pyrethroid cypermethrin at concentrations found in the environment and their potential deleterious effects on nontarget aquatic organisms.

Protective efficacy of naringenin against cadmium-induced redox imbalance in Labeo rohita: an integrated biomarker approach.

The protective efficacy of dietary naringenin (NG) has been investigated against the toxicity caused by cadmium chloride (CdCl2) using biomarkers of oxidative stress in the liver, gills and kidney of Labeo rohita. The fish were exposed to environmentally relevant concentrations of CdCl2 (0.37 and 0.62 mg/L) and simultaneously orally administered with NG (50 mg/kg bw/day) for 60 days. Tissue (gills, liver and kidney) samples were collected on days 15, 30 and 60 of the experiment and analysed for endogenous antioxidants and oxidative stress biomarkers. CdCl2 exposure for 15 and 30 days induced the development of adaptive mechanism as demonstrated by the enhanced activities of superoxide dismutase, catalase, glutathione peroxidase and glutathione-S-transferase in all three tissues. However, on the 60th day, CdCl2-induced oxidative damage was stipulated by a decline in the enzyme activities and reduced glutathione (GSH) content significantly (p < 0.05) below control levels along with enhanced levels of lipid peroxidation. Oral administration of NG in toxicant exposed fish significantly restored the altered levels of antioxidants, oxidative enzymes and lipid peroxidation. Besides, integrated biomarker response (IBR) analysis was applied by combining all the biomarkers to indicate the overall stress response index. IBR analysis confirmed the altered levels of biomarkers, the oxidative stress induced by CdCl2 exposure and the ameliorative potential of NG. The present study suggested that NG might have protective role against Cd-induced oxidative insult which might be ascribed to the ability of NG to chelate metals and scavenge free radicals.

Oxidative Stress and Toxico-Pathic Branchial Lesions in Cyprinus carpio Exposed to Malachite Green.

Gill is the frontier tissue to come in direct contact with aquatic toxicants. Malachite green (MG) commercial textile dye was assessed for its impact on the gill cytoarchitecture. Cyprinus carpio were exposed to 0.087 and 0.146 mg/L of MG for 60 days. The tissue was processed, and HE stained slides revealed histo-pathic lesions such as lamellar curling, edema, necrosis, telangiectasia, aneurysm, and vacuolization. Scanning electron microscopy reported aberrations in lamellae and microridges of the epithelium. At the cellular level, transmission electron microscopy exhibited nuclear alterations in form of pyknosis and mitochondrial swelling followed by cristolysis. Pillar cells displayed cytoplasmic vacuolization and leukocyte infiltration, and goblet cell containing varied shaped and density mucous globules. The biochemical analysis supported the ultrastructural alterations and showed a negative impact of MG on the antioxidative enzymes (CAT, SOD, GSH), while levels of MDA were found to be significantly elevated. Thereby, concluding MG induced branchial toxicity in the fish.

Impact of cypermethrin in nephrocytes of freshwater fish Catla catla.

The kidney of Catla catla, chronically exposed to sub-lethal concentrations (0.24 µg/L and 0.41 µg/L) of cypermethrin revealed a significant elevation in the activity of antioxidants superoxide dismutase (SOD), catalase (CAT), glutathione-s-transferase (GST) and reduced glutathione (GSH) after 15 days, followed by a decline of up to 45 days. Lipid peroxidation (LPO) remained elevated throughout the exposure duration. Histology presented proliferated haematopoietic tissue, tubular and glomerular degeneration. The maximum increase in the mean degree of tissue change (DTC) was observed on the 45th day of treatment. Ultra-structure study depicted cytoplasmic vacuolation, fragmented RER, the proliferation of lysosomes, mitochondrial degeneration, and degenerative changes in the epithelial lining of renal tubules. Principal component analysis (PCA) of various biomarkers generated two components PCI (SOD, GST, GSH, LPO and DTC) and PCII (CAT). These findings suggest that long term exposure to cypermethrin can lead to various pathological alterations in the fish kidney which in turn might interfere with normal renal excretory mechanism.

Cassia fistula ameliorates chronic toxicity of cypermethrin in Catla catla.

Protective effects of Cassia fistula, in Catla catla exposed to synthetic pyrethroid cypermethrin were evaluated. Fish, after chronic exposure to environmentally relevant sub-lethal concentration 0.41 µg/l of the pesticide were assessed for antioxidant activity, histopathological and ultrastructural alterations. Significant (p < 0.05) decrease in the activities of antioxidants such as CAT, SOD, GST, GSH was registered, whereas LPO level got elevated. Histological damage depicted necrosis, epithelial hypertrophy, hyperplasia and fusion of secondary lamellae and changes in gill vasculature. Histopathological alteration index was employed for the semi quantitative evaluation of the degree of tissue change (DTC). Transmission electron microscopy displayed swollen and distorted mitochondria, damaged chloride cells and necrosis. Dietary supplementation of Cassia fistula bark extract significantly (p < 0.05) improved the antioxidant activity, reduced lipid peroxidation and prevented histopathological alterations. The findings suggest that sub-lethal concentration of cypermethrin is toxic to fish. The study also draws attention towards potential of plant derived antioxidants in mitigating pesticide induced toxic effects.

Nephroprotective effect of Emblica officinalis fruit extract against malachite green toxicity in piscine model: Ultrastructure and oxidative stress study.

Malachite green (MG) is a multi-application dye with raised concern as aquatic toxicant. Cyprinus carpio fingerlings were exposed to MG and simultaneously fed with Emblica officinalis (EO) fruit extract to mitigate the MG induced nephrotoxicity. MG exposure developed depressed activity of antioxidant enzymes such as catalase, superoxide dismutase, glutathione-s-transferase, and reduced glutathione, while levels of malondialdehyde got significantly (p < .05) elevated after 60 days MG exposure. H&E staining revealed increased intracellular space, vacuolation, fluid-filled spaces, damaged brush border lining of proximal convoluted tubule (PCT), focal necrosis, nuclear transformations and shrunken glomerulus. Further, Transmission electron microscope study detailed structural cytotoxicity with the appearance of the heterochromatic nucleus, electron-dense lysosomal bodies, damaged brush border lining of PCT, and necrosis; glomerulus exhibited erythrocyte infiltration and fused pedicels of podocyte. While, EO extract supplemented diet culminated in the restoration of the renal cytoarchitecture and significantly (p < .05) enhanced activity of antioxidant enzymes.

Ameliorating effect of ascorbic acid on fenvalerate induced ultrastructural changes in scales, erythrocytes and gills of Ctenopharyngodon idella (Valenciennes, 1844).

Fenvalerate (type II synthetic pyrethroid), widely used in agricultural practices, find its way into aquatic ecosystem through air, by runoff, or by percolation to groundwater. It is an extremely toxic insecticide for aquatic organisms especially fish. In the present study, the fenvalerate (FEN) induced toxicity and the protective efficacy of ascorbic acid (AA) against FEN in Ctenopharyngodon idella was evaluated by studying the structural alterations in scales, erythrocytes and gills. The fishes were exposed to 1.2 µg/L and 2 µg/L of FEN and orally administered with 1000 mg/kg diet of AA. The fishes were scrutinized on 15th, 30th and 60th day of experiment. Scanning electron microscopic studies (SEM) of FEN-treated fish revealed extensive morphological alterations on the microstructure of scales including deformed focus, uprooted lepidonts and tubercles, hole formation and worn out calcareous material from the surface. FEN intoxication induced severe damage on erythrocytes including formation of dacrocytes, serrated spherocytes, echinocytes with oozed out cytoplasmic content, contracted plasma membrane and appearance of lobopodial projections. Ultrastructural studies in gills declared profound lesions in the form of aneurysm, loss of secondary lamellae and destructed microstructures of pavement cells. On the other hand, supplementation of AA in diet mitigated the impairment provoked by FEN on the scales, erythrocytes and gills due to its antioxidant properties.

Assessment of cypermethrin induced hepatic toxicity in Catla catla: A multiple biomarker approach.

The study was designed to evaluate chronic toxicity of pyrethroid pesticide cypermethrin through biochemical, histopathological, ultrastructural and molecular biomarkers in liver of freshwater carp Catla catla. The fish were exposed to two sub-lethal concentrations (0.21 µg/L and 0.41 µg/L) for a period of 45 days. Compared to the control, a significant (p < 0.05) increase in the activity of enzymatic antioxidants catalase (CAT), superoxide dismutase (SOD) and glutathione-S-transferase (GST), and glutathione content (GSH) was registered after initial 15 days of exposure to the toxicant, followed by decline on 30th and 45th day. Whereas, MDA level remained elevated throughout the exposure duration at both the tested concentrations. Light microscopy revealed changes like sinusoidal dilation, vacuolation, pycnosis, karyolysis, nuclear pleomorphism, lymphocyte infiltration in liver of the exposed fish, with highest mean degree of tissue change (DTC) value of 58.6 ± 3.19 on 45th day. Ultrastructurally, cytopasmic vacuolation, reduced glycogen granules, dilated RER, deformed nuclear membrane, swollen & distorted mitochondria and augmentation in lipid bodies were the prominently observed cytopathological alterations. These anomalies increased in time-concentration dependent manner, being most severe after 45 days at higher concentration. The gene expression levels of Gadd-45α and Bcl-2 depicted altered patterns. Gadd-45α exhibited significant upregulation by 45th day, while Bcl-2 demonstrated initial upregulation, with subsequent downregulation on 30th and 45th day. Principal component analysis (PCA) generated two components, PC1 (SOD, GSH, MDA and DTC) and PC2 (CAT and GST). The findings suggest that cypermethrin inflicts marked hepatototoxic effects on Catla catla even at sub-lethal concentrations.

Genotoxicity induced by hexavalent chromium leading to eryptosis in Ctenopharyngodon idellus.

The initiation of eryptosis as a result of genotoxic action of Cr(VI), seen through micronucleus and comet assay in the peripheral erythrocytes of Ctenopharyngodon idellus was evaluated through RT-qPCR. For this, fish was exposed to sublethal concentration of hexavalent chromium (5.30 and 10.63 mg/L), and the blood was sampled on different endpoints (15, 30 and 45 days). Accumulation of chromium in the erythrocytes was also studied, which depicted a significant increase in toxicant concentration and time dependent manner. Both concentrations of hexavalent chromium induced DNA damage, visible in the form of comet tails. The presence of micronuclei in the erythrocytes was accompanied with occurrence of nuclear bud (NBu), lobed nucleus (Lb), notched nucleus (Nt), vacuolated nucleus (Vn), binucleated cell (Bn) as nuclear abnormalities; and acanthocytes (Ac), echinocytes (Ec), notched cells (Nc), microcytes (Mc) and vacuolated cytoplasm (Vc) as cytoplasmic abnormalities. The expression of genes related to intrinsic apoptotic pathway induced by Cr(VI) presented significant (p < 0.05) upregulation in the expression of p53, Bax, Apaf-1, caspase9 and caspase3, and downregulation of Bcl2; inferring the initiation of apoptotic pathway. The ration of Bax and Bcl2 also appended the apoptotic state of the erythrocytes. From the present investigation, it can be concluded that genotoxicity induced by hexavalent chromium lead to eryptosis in C. idellus.

Hexavalent chromium-induced toxic effects on the antioxidant levels, histopathological alterations and expression of Nrf2 and MT2 genes in the branchial tissue of Ctenopharyngodon idellus.

Ability of hexavalent chromium to accumulate and induce oxidative stress has been studied in the gills of Ctenopharyngodon idellus, with the resulting damage in the form of altered endogenous antioxidant enzyme activity and, histopathology in the tissue. The fish were exposed to 5.3 (C1) and 10.63 mg/L (C2) of hexavalent chromium and were scrutinised on 15th, 30th and 45th day of toxicant exposure. Oxidative stress studied in terms of lipid peroxidation and glutathione levels and the antioxidant enzymes activity also exhibited alterations. The histopathological modifications in gills announced lesions in the form of hyperplasia, aneurysm, lamellar fusion, focal proliferation, epithelial degeneration and necrosis with loss of lamellae, bringing irreversible damage on 45th day with mean degree of tissue change value of 100.35 ±â€¯10.69. Bioaccumulation of chromium, and increased anomalies in branchial tissue exhibited damage in concentration and time-dependent manner. The ultrastructural anomalies in the cellular morphology in the epithelial cells of filaments and lamellae, exhibited pleomorphic nuclei, swollen mitochondria, extensive vacuolation and loss of microridges in pavement cells. The tissue also displayed altered regulation of Nrf2 and Mt2 following Cr(VI) exposure with maximum downregulation on 45th day by 61 and 53%, respectively. PCA generated two principal components, PC1 (GSH, GST, CAT and SOD) and PC2 (DTC, MDA and Cr(VI) concentration). Thus, it can be concluded that accumulation of Cr(VI) induces alteration in the gene expression of Nrf2 and Mt2 leading to the development of oxidative stress, ensuing various pathological changes creating hindrance in fish survival.

Evaluating the protective efficacy of Silybum marianum against deltamethrin induced hepatotoxicity in piscine model.

With the known facts of deltamethrin toxicity in aquaculture, efforts have been made to ameliorate its toxicity with supplementation of Silybum marianum plant extract in Cyprinus carpio. For validating its efficacy, standard techniques of histopathology, anti-oxidant profile and RAPD-PCR were used. By performing acute toxicity bioassay, 96 h LC50 of the pesticide to C. carpio was determined and was found to be 2 µg/L. Histopathological alterations were comprised of nuclear alterations, vacuolisation and oedema in the hepatocytes. Chronic exposure to the toxicant induced significant changes in antioxidant defense system (CAT, SOD, GSH and GST levels), lipid peroxidation being prominent. Diet supplementation with silymarin appeared to modulate the oxidative stress, histopathological alterations and genotoxic damage caused by the pesticide in the fish. RAPD-PCR studies revealed deltamethrin induced toxicity and its effective amelioration in form of restoration of bands which were lost in toxicant exposed DNA profile.

Malachite Green Induced Ultrastructural Corneal Lesions in Cyprinus carpio and Its Amelioration Using Emblica officinalis.

Malachite green, a multi-purpose dye induces cyto-toxicity upon its entry and bioaccumulation in tissues. A semi-static chronic (60 days) bioassay was conducted by exposing Cyprinus carpio to sublethal concentration of the dye and Emblica officinalis in four experimental groups viz control, malachite green, E. officinalis, and malachite green + E. officinalis. Effect of dye on the cornea was investigated considering ultra-structural alterations owing to its direct contact to the pollutant in the aquatic medium. SEM studies on corneal epithelium revealed broken continuity of pavement cells, shrunk microplicae, increased intra-microplicae distance, globularization and epithelial uplifting, thereby affecting the integrity of corneal surface and tear film adherence. Whereas dietary supplementation with the plant extract served to restore cytoarchitecture with appearance of large number of regenerating cells. Both lesions and restoration were found to be duration dependent. Thus, E. officinalis can be considered as an effective ameliorant against malachite green induced toxicity.

Toxicopathic Branchial Lesions in Grass Carp (Ctenopharyngodon idellus) Exposed to Chlorpyrifos.

Present investigation aimed to assess the alterations in branchial architecture of Ctenopharyngodon idellus exposed to chlorpyrifos. 96 h LC50 of chlorpyrifos to C. idellus was found to be 7.24 µg/L. Fish were exposed chronically to 2.41 and 1.44 µg/L sublethal concentrations (ecologically relevant concentrations) of chlorpyrifos for 15, 30 and 60 days. The gills of the CPF exposed fish showed partially affected secondary lamellae with shortening and curling, increased inter-lamellar space, sloughed off epithelium, hyperplasia and necrosis of pavement cells. On 60 days exposure, more pronounced degenerative changes, including severe necrosis, hyperplasia and hypertrophy of pavement cells, eroded and uplifted lamellar epithelium were noticed. Ultrastructural alterations include distorted nucleus and mitochondria in pavement cells, marked contraction in cisternae of golgi complex. Particularly in chloride cells, abundant mitochondria with distorted shape and size, dilation in between their cristae and tubular network were observed. The severity of damage was more profound at 60 days exposure. Thus, the observed cytopathological lesions of fish gills can be used as sensitive biomarkers for pesticide contamination.

In vivo genotoxicity and cytotoxicity assessment of cadmium chloride in peripheral erythrocytes of Labeo rohita (Hamilton).

Cadmium chloride (CdCl2) induced genotoxicity and cytotoxicity has been assessed in the peripheral blood erythrocytes of freshwater fish Labeo rohita exposed to 0.37 and 0.62mg/L of CdCl2 in water for 100 days. The blood samples of the fish were collected at different intervals (days 1, 3, 5, 10, 15, 30, 60 and 100) of exposure period to analyze DNA damage using comet assay and the occurrence of micronuclei and other cellular anomalies. The results of comet assay showed a significant increase in the mean percentage of tail DNA at both the concentrations. Exposure to CdCl2 also induced micronuclei in addition to many nuclear abnormalities such as nuclear bud, binucleates, lobed, notched and vacuolated nuclei. Cytoplasmic abnormalities like echinocytes, acanthocytes, notched, microcytes and cells with vacuolated cytoplasm were also observed. The metal exposed groups showed significant variation in the frequency of cellular abnormalities as well as the extent of DNA damage in comparison to controls. These frequencies increased significantly (p<0.05) in concentration dependent manner, peaking on 10th day while a decreasing trend was observed after 15 days of the exposure period.

The truth about medical negligence.

A vast majority of people believes that doctors are a negligent lot. This often implies that doctors are not courteous enough, are brusque to the point of being rude, are not available when needed, and prescribe unnecessary laboratory tests, scans and medicines. At a function organised by the Indian Medical Association on medical negligence, a veteran politician, otherwise very articulate, was at a loss to define it. Finally, he said, "Don't you people leave your scissors or gauze pieces in the stomachs of patients?" The usual complaints of being rude and unsympathetic pertain more to a lack of professional courtesy, which is our national trait. Doctors are no exception, though this national trait only explains their conduct and does not excuse or justify it.

Intravesical migration of intrauterine contraceptive devices with stone formation.

Intrauterine contraceptive devices (IUCD) have been associated with the multitude of complications. We present a rare case report of a 30-year-old female in whom the IUCD (Cu-T) migrated into the urinary bladder leading to calculus formation. The migrated IUCD encrusted with stones was successfully retrieved.